Standard Operating Procedure

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1 Subject Specimens for Anaereobic Culture, Collection of Index Number Lab-1217 Section Laboratory Subsection Specimen Collection and Processing Category Departmental Contact Sarah Stoner Last Revised 8/8/2017 References Required document for Laboratory Accreditation by the College of American Pathologists (CAP), Centers for Medicare and Medicaid Services (CMS) and or COLA. Applicable To Employees of Gundersen Health System clinical laboratories, Gundersen St. Joseph s Health Services laboratories, Gundersen Tri-County Memorial laboratories, Gundersen Palmer Lutheran Hospital Laboratory and Gundersen Boscobel Area Health Services. Detail PRINCIPLE: A crucial factor affecting the ultimate success of anaerobic cultures is the transport of specimens. The bacteria must be protected from the lethal effects of oxygen during from time of collection of the specimen until it is set up anaerobically in the laboratory. Specimens should be transported and processed rapidly to avoid loss of fastidious oxygen sensitive anaerobes and to prevent overgrowth of facultative bacteria. Since anaerobes may cause or contribute to infections of all types, it is clear that all specimens which are free of "contamination" with normal flora should be cultured anaerobically. Proper collection, with care to avoid inclusion of normal flora cannot be overemphasized. Indigenous anaerobes are often present in such large numbers that even minimal contamination of a specimen with normal flora can give very misleading results. The BD Liquid Amies Elution Swab (ESwab) Collection and Transport system is intended for the collection and transport of swab samples. It incorporates a modified Liquid Amies transporting medium, which can sustain the viability of clinically important aerobes, anaerobes, and fastidious bacteria. The ESwab system is a maintenance medium comprised of inorganic phosphate buffer, calcium and magnesium salts, and sodium chloride with a reduced environment due to the presence of sodium thioglycollate. CLINICAL SIGNIFICANCE: N/A SPECIMEN: Site Acceptable Specimens Unacceptable Specimens Head and Neck 1. Abscess aspirate obtained by needle and syringe after surface decontamination. 3. Surgically obtained ESwab when aspiration is not feasible. 1. Throat or nasopharyngeal swabs 2. Gingival swabs 3. Superficial material collected with swabs. Page 1 of 5

2 Lungs 1. Transtracheal aspirate 2. Materials from percutaneous lung puncture. 3. Biopsy material surgically obtained. 4. Bronchoscopic specimen obtained by 1. Expectorated sputum 2. Induced sputum 3. Endotracheal aspirate 4. Bronchoscopic specimens not specially collected. protected brush. 5. Thoracotomy specimen 6. Surgically obtained ESwab specimen. Central Nervous System 1. Abscess aspirate obtained by needle or syringe. 3. Surgically obtained ESwab. Abdomen 1. Peritoneal fluid obtained by needle and syringe 2. Bile 3. Biopsy material surgically obtained. 4. Surgically obtained ESwab. Urinary tract Suprapubic aspirate 1. Voided urine Female genital tract Bone and joint Soft tissue Stomach and small bowel 1. Culdoscopy specimens 2. Endometrial aspirate obtained by suction or protected collector. 3. Abscess aspirate obtained by needle and syringe. 4. Biopsy material surgically obtained 5. Surgically obtained ESwab 6. IUD for Actinomyces sp. 1. Aspirate obtained by needle and syringe 3. Surgically obtained ESwab. 1.Aspirate by needle and syringe 2. Aspirate from sinus tract obtained by needle and small plastic catheter 3. Deep aspirate of open-wound margin obtained through decontaminated skin 4. Deep aspirate of surface ulcer obtained through decontaminated skin 5. Surgically obtained deep ESwab specimen Only for workup of blind-loop or malabsorption syndrome. REAGENTS AND MATERIALS: BD Liquid Amies Elution Swab (ESwab) Collection and Transport system. Sterile needle and syringe. Sterile screw top container. BD Plain Red Top (preferred), Citrate, or EDTA Vacutainer tubes. 2. Catheterized urine Vaginal or cervical swabs Superficial material collected with swabs Superficial material collected from skin surface or edges of wound Page 2 of 5

3 BD Standard Aerobic and Anaerobic Lytic Blood culture bottles EQUIPMENT/INSTRUMENTATION: N/A QUALITY CONTROL: Do not use BD Liquid Amies Elution Swab (ESwab) Collection and Transport system or other collection and transport devices if there is evidence of damage or contamination to the product, leakage, if the product is expired, the collection kit is opened, or has other signs of deterioration. Each lot is tested by the manufacturer for appropriate performance standards. Implementation CHOICE OF SPECIMEN A. Tissue - 1st choice for best recovery 1. Tissue specimens may more accurately indicate the bacteriology of the infectious process, and anaerobic bacteria survive more readily in this type of specimen. 2. Tissue should be submitted in a sterile screw capped cup. 3. Do not place in fixative or preservative. B. Fluids - 2nd choice of specimen type 1. Draw fluid into syringe and expel excess air. a. For large volume fluids (>10mL), sterilize the rubber stopper of the aerobic and anaerobic blood culture bottles (large volume fluids) with alcohol prep pad, let dry, and inject fluid. Send specimen to the lab as soon as possible. b. For smaller volume fluids (<10mL), sterilize the rubber stopper of a vacutainer with alcohol prep pad, let dry, and inject fluid. DO NOT REMOVE CAP OF VACUTAINER AND ADD FLUID. Send specimen to the lab as soon as possible. 2. If no vial is available, the plastic syringe, with a sterile stopper replacing the needle tip, may be sent to the lab. However, the culture must be set up within twenty minutes. C. Swab Cultures - 3rd choice, least desirable for best recovery. 1. Once a specimen is collected, it should be placed immediately in contact with the transport medium. ESwabs should be transported directly to the laboratory, preferably within 2 hrs of collection to maintain optimum organism viability. If immediate delivery or processing is delayed, then specimens should be stored at 4-8C or room temperature (22-25C) and processed within 48hrs. PROCEDURAL NOTES A. CLUES TO ANAEROBIC INFECTIONS 1. Foul-smelling discharge. 2. Location of infection in proximity to a mucosal surface. 3. Necrotic tissue, gangrene, pseudomembrane formation. 4. Gas in tissues or discharges. 5. Endocarditis with negative routine blood cultures. 6. Infection associated with malignancy or other process resulting in tissue destruction. 7. Infection related to the use of aminoglycocides (oral, parenteral, or topical). 8. Septic thrombophlebitis. 9. Infection following human or other bites. Page 3 of 5

4 10. Black discoloration of blood-containing exudates; these exudates may fluoresce red under ultraviolet light. 11. Presence of sulfur granules in discharges (actinomycosis). 12. Clinical setting suggestive of anaerobic infection. 13. Classical clinical picture. B. UNACCEPTABLE SPECIMENS Criteria for Rejection of Unacceptable Specimens for Anaerobe Culture: Category Criteria for Rejection Action Identification Discrepancy between patient identification on request form and on specimen container. No identification on container. Contact sender to correct identification, and then fill out an incident report. DO NOT process. Try to determine sending location and notify of recollection needed. Specimen source or type of culture ordered Call for necessary information not on request form. Specimen Specimen container broken, spilled or Request new specimen. leaking. Swab specimen received in a culturette. Request new specimen. Voided urine. Request a suprapubic bladder aspiration. Feces A regular culture not done. Normal flora of Feces contain from species of anaerobes at one time. We will screen for clostridium difficile only. Sputum Request transtracheal aspirate to avoid normal flora. Freshly ruptured appendix Culture will be contaminated with normal fecal flora screen for anaerobes only and call doctor. Throat culture Culture not done. Request sampling of the abscess while eliminating the normal flora. *Before rejected specimens are discarded, make sure that a second specimen is or can be obtained. CALCULATIONS: N/A INTERPRETATION: N/A LIMITATIONS: N/A REVIEW AND CHANGES: This document and all attached forms should be reviewed optimally on an annual basis, with 2 years as the maximum review date. Review will be done by the Technical Leader, Supervisor, Manager, Medical Page 4 of 5

5 Director or designated person. Changes require retyping document or form and review by the Medical Director. REFERENCES: 1. Anaerobic Bacteria in Human Disease, Finegold, Sydney M., M.D., Academic Press, New York, ESwab package insert, BD Manual of Clinical Microbiology, Jorgensen, J. H., et al. 11 th ed. ASM Press, 2015, pp Essential Procedures for Clinical Microbiology, Henry D. Isenberg, ASM Press, 1998, pp (Formerly Anaerobic Micro policy Lab-7170) Page 5 of 5